Method for administering insulinotropic peptides

ABSTRACT

The claimed invention relates to a method of administering glucagon-like peptide-1 molecules by inhalation, a method for treating diabetes by administering glucagon-like peptide-1 molecules by inhalation, and a method for treating hyperglycemia by administering glucagon-like peptide-1 molecules by inhalation.

CROSS-REFERENCE

[0001] This application claims the benefit of U.S. ProvisionalApplication No. 60/098,273, filed Aug. 28, 1998, and U.S. ProvisionalApplication No. 60/100,012, filed Sep. 11, 1998.

FIELD OF THE INVENTION

[0002] This invention relates to methods of treating humans sufferingfrom diabetes and insulin resistance. In particular, the inventionrelates to the pulmonary delivery of glucagon-like peptide-1 (GLP-1) andanalogs thereof for systemic absorption through the lungs to eliminatethe need for administering anti-diabetic compounds by injection.

BACKGROUND OF THE INVENTION

[0003] Glucagon-like peptide-1 was first identified in 1987 as aincretin hormone, a peptide secreted by the gut upon ingestion of food.Glucagon-like peptide-1 is secreted by the L-cells of the intestineafter being proteolytically processed from the 160 amino acid precursorprotein, preproglucagon. Cleavage of preproglucagon first yieldsglucagon-like peptide-1, a 37 amino acid peptide that is poorly active.A subsequent cleavage of the peptide bond between residues 6 and 7yields biologically active glucagon-like peptide-1 referred to as GLP-1(7-37). It should be noted that this specification uses the nomenclaturescheme that has developed around this hormone. By convention in the art,the amino terminus of GLP-1(7-37) has been assigned number 7 and thecarboxy terminus number 37. Approximately 80% of the GLP-[(7-37) that issynthesized is amidated at the C-terminal after removal of the terminalglycine residue in the L-cells. The biological effects and metabolicturnover of the free acid GLP-1 (7-37), and the amide, GLP-1 (7-36)NH₂,are indistinguishable. As used herein, these two naturally-occurringforms will be referred to collectively as GLP-1.

[0004] GLP-1 is known to stimulate insulin secretion (insulinotropicaction) causing glucose uptake by cells which decreases serum glucoselevels (see, e g., Mojsov, S., Int. J. Peptide Protein Research,40:333-343 (1992)). Numerous GLP-1 analogs and derivatives demonstratinginsulinotropic action are known in the art. Also it has beendemonstrated that the N-terminal histidine residue (His 7) is veryimportant to insulinotropic activity of GLP-1 (Suzuki, S., et al.Diabetes Res.; Clinical Practice 5 (Supp. 1):S30 (1988).

[0005] Multiple authors have demonstrated the nexus between laboratoryexperimentation and mammalian, particularly human, insulinotropicresponses to exogenous administration of GLP-1. See, e.g., Nauck, M. A.,et al., Diabetologia, 36:741-744 (1993); Gutniak, M., et al., NewEngland J. of Medicine, 326(20):1316-1322 (1992); Nauck, M. A., et al.,J. Clin. Invest., 91:301-307 (1993); and Thorens, B., et al., Diabetes,42:1219-1225 (1993)].

[0006] GLP-1 based peptides hold great promise as alternatives toinsulin therapy for patients with diabetes who have failed onsulfonylureas. GLP-1 has been studied intensively by academicinvestigators, and this research has established the following forpatients with type II diabetes who have failed on sulfonylureas:

[0007] 1) GLP-1 stimulates insulin secretion, but only during periods ofhyperglycemia. The safety of GLP-1 compared to insulin is enhanced bythis property of GLP-1 and by the observation that the amount of insulinsecreted is proportional to the magnitude of the hyperglycemia. Inaddition, GLP-1 therapy will result in pancreatic release of insulin andfirst-pass insulin action at the liver. This results in lowercirculating levels of insulin in the periphery compared to subcutaneousinsulin injections.

[0008] 2) GLP-1 suppresses glucagon secretion, and this, in addition tothe delivery of insulin via the portal vein helps suppress the excessivehepatic glucose output in diabetic patients.

[0009] 3) GLP-1 slows gastric emptying which is desirable in that itspreads nutrient absorption over a longer time period, decreasing thepostprandial glucose peak.

[0010] 4) Several reports have suggested that GLP-1 may enhance insulinsensitivity in peripheral tissues such as muscle and fat.

[0011] 5) Finally, GLP-1 has been shown to be a potential regulator ofappetite.

[0012] Meal-time use of GLP-1 based peptides offers several advantagesover insulin therapy. Insulin therapy requires blood glucose monitoring,which is both expensive and painful. The glucose-dependency of GLP-1provides an enhanced therapeutic window in comparison to insulin, andshould minimize the need to monitor blood glucose. Weight gain also canbe a problem with intensive insulin therapy, particularly in the obesetype II diabetic patients.

[0013] The therapeutic potential for native GLP-1 is further increasedif one considers its use in patients with type I diabetes. A number ofstudies have demonstrated the effectiveness of native GLP-1 in thetreatment of insulin dependent diabetes mellitus. Similar to patientswith type II diabetes, GLP-1 is effective in reducing fastinghyperglycemia through its glucagonostatic properties. Additional studieshave indicated that GLP-1 also reduces postprandial glycemic excursionsin type I patients, most likely through a delay in gastric emptying.These observations indicate that GLP-1 may be useful as a treatment intype I and type II patients.

[0014] To date administration of clinically proven peptide hormones andas well as GLP-1 has generally been accomplished by subcutaneousinjection which-is both inconvenient and unattractive. Therefore, manyinvestigators have studied alternate routes for administering peptidehormones such as oral, rectal, transdermal, and nasal routes. Thus far,however, these routes of administration have not resulted in clinicallyproven peptide hormone therapy.

[0015] It has been known for a number of years that some proteins can beabsorbed from the lung. For example, insulin administered by inhalationaerosol to the lung was first reported by Gaensslen in 1925. Despite thefact that a number of human and animal studies have shown that someinsulin formulations can be absorbed through the lungs, pulmonarydelivery of peptide hormones has not been vigorously pursued because ofvery low bioavailability. Larger proteins, such as cytokines and growthfactors which are generally larger than 150 amino acid residues, areoften readily absorbed by the cells lining the alveolar regions of thelung. Pulmonary absorption of smaller proteins is however much lesspredictable; though insulin (51 residues), calcitonin (32 residues) andparathyroid hormone (34 residues) have been reported to be systemicallyabsorbed through the pulmonary route. See U.S. Pat. No. 5,607,915,herein incorporated by reference. Despite systemic absorption by thelung of some small protein hormones, the pharmacodynamics associatedwith pulmonary delivery of peptides is unpredictable.

[0016] Thus, there is a need to provide a reliable pulmonary method ofdelivering GLP-1 and related analogs because it would offer patients anattractive, non-invasive alternative to insulin. This need isparticularly true since insulin has a very narrow therapeutic indexwhile GLP-1 treatment offers a way to normalize blood glucose only inresponse to hyperglycemic conditions without the threat of hypoglycemia.

[0017] Not all protein hormones can be efficiently absorbed through thelungs, and there are many factors that affect it. Absorption of proteinsin the lung is largely dependent on the physical characteristics of theprotein. Thus, even though pulmonary delivery of some protein hormoneshas been observed, the physical properties and short length of GLP-1 andsome related peptides made it unclear whether such peptides could beeffectively delivered through the pulmonary route.

[0018] Efficient pulmonary delivery is dependent on the ability todeliver the protein to the deep lung alveolar epithelium. Proteinparticles that lodge in the upper airway epithelium are not absorbed toa significant extent because the overlying mucus functions to trap, andthen clear debris by mucociliary transport up the airway. This mechanismis also a major contributor to low bioavailability. The extent to whichproteins are not absorbed and instead eliminated by these routes dependson their solubility, their size, and other largely uncharacterizedmechanisms.

[0019] Even when a peptide hormone can be reproducibly delivered to thedeep lung alveolar epithelium, it is difficult to predict whether itwill be rapidly absorbed and transported to the blood. Absorption valuesfor some proteins delivered through the lungs have been calculated andrange from fifteen minutes for parathyroid hormone (1-34) to 48 hoursfor glycosylated α1-antitrypsin. Moreover a variety of endogenouspeptidases exist in the lung which can degrade peptides prior toabsorption. Thus, the longer it takes for a peptide particle to dissolveand be absorbed, the greater the chance for enzymatic inactivation.Thus, because of the small size of GLP-1 and its inherent susceptibilityto certain enzymes, it was most surprising to find that an aerosolizedGLP-1 analog could be reproducibly and effectively delivered through thelungs.

SUMMARY OF THE INVENTION

[0020] The present invention relates to a method for administering aglucagon-like peptide-1 molecule comprising, administering an effectiveamount of the peptide to a patient in need thereof by pulmonarydelivery. The present invention also relates to a method for treatingdiabetes comprising, administering an effective dose of a glucagon-likepeptide-1 to a patient in need thereof by pulmonary delivery. Anotheraspect of the invention relates to a method for treating hyperglycemiacomprising, administering an effective dose of a glucagon-like peptide-1to a patient in need thereof by pulmonary delivery. Preferably, theglucagon-like peptide-1 molecule is delivered by inhalation and to thelower airway of the patient.

[0021] The glucagon-like peptide-1 can be delivered in a carrier, as asolution or suspension, or as a dry powder, using any of a variety ofdevices suitable for administration by inhalation. Preferably, theglucagon-like peptide-1 is delivered in a particle size effective forreaching the lower airways of the lung.

DETAILED DESCRIPTION OF THE INVENTION

[0022] The term “GLP-1” refers to human glucagon-like peptide-1 whosesequences and structures are known in the art. See U.S. Pat. No.5,120,712, herein incorporated by reference. As previously discussed,there are two native forms of human GLP-1, GLP-1 (7-37) and GLP-1(7-36)NH₂ which will be distinguished only when necessary.

[0023] The term “GLP-1 analog” is defined as a molecule having one ormore amino acid substitutions, deletions, inversions, or additionscompared with GLP-1. Many GLP-1 analogs are known in the art andinclude, for example, GLP-1(7-34) and GLP-1 (7-35), GLP-1 (7-36),Val⁸-GLP-1 (7-37), Gln⁹-GLP-1 (7-37), D-Gln⁹-GLP-1 (7-37),Thr¹⁶-Lys¹⁸-GLP-1 (7-37), and Lys¹⁸-GLP-1 (7-37). Preferred GLP-1analogs are GLP-1 (7-34) and GLP-1 (7-35), which are disclosed in U.S.Pat. No. 5,118,666, herein incorporated by reference.

[0024] The term “GLP-1 derivative” is defined as a molecule having theamino acid sequence of GLP-1 or a GLP-1 analog, but additionally havingchemical modification of one or more of its amino acid side groups,a-carbon atoms, terminal amino group, or terminal carboxylic-acid group.A chemical modification includes, but is not limited to, adding chemicalmoieties, creating new bonds, and removing chemical moieties.Modifications at amino acid side groups include, without limitation,acylation of lysine e-amino groups, N-alkylation of arginine, histidine,or lysine, alkylation of glutamic or aspartic carboxylic acid groups,and deamidation of glutamine or asparagine. Modifications of theterminal amino-include, without limitation, the des-amino, N-loweralkyl, N-di-lower alkyl, and N-acyl modifications. Modifications of theterminal carboxy group include, without limitation, the amide, loweralkyl amide, dialkyl amide, and lower alkyl ester modifications. Loweralkyl is C₁-C₄ alkyl. Furthermore, one or more side groups, or terminalgroups, may be protected by protective groups known to theordinarily-skilled protein chemist. The a-carbon of an amino acid may bemono- or di-methylated.

[0025] The term “GLP-1 molecule” means GLP-1, GLP-1 analog, or GLP-1derivative.

[0026] Another preferred group of GLP-1 analogs is defined by theformula:

R₁-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Y-Gly-Gln-Ala-Ala-Lys-Z-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-R₂  (SEQ.ID NO:1)

[0027] and the pharmaceutically-acceptable salts thereof, wherein: R₁ isselected from the group consisting of L-histidine, D-histidine,desamino-histidine, 2-amino-histidine, b-hydroxy-histidine,homohistidine, alpha-fluoromethyl-histidine, and alpha-methyl-histidine;X is selected from the group consisting of Ala, Gly, Val, Thr, Ile, andalpha-methyl-Ala; Y is selected from the group consisting of Glu, Gln,Ala, Thr, Ser, and Gly; Z is selected from the group consisting of Glu,Gln, Ala, Thr, Ser, and Gly; and R₂ is selected from the groupconsisting of NH₂, and Gly-OH; providing that when R₁ is His, X is Ala,Y is Glu, and Z is Glu, R₂ must be NH₂.

[0028] Yet another preferred group of compounds consistent with thepresent invention is disclosed in WO 91/11457 (U.S. Pat. No. 5,545,618,herein incorporated by reference) and consists essentially of GLP-1(7-34), GLP-1 (7-35), GLP-1 (7-36), or GLP-1 (7-37), or the amide formsthereof, and pharmaceutically-acceptable salts thereof, having at leastone modification selected from the group consisting of:

[0029] (a) substitution of glycine, serine, cysteine, threonine,asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine,methionine, phenylalanine, arginine, or D-lysine for lysine at position26 and/or position 34; or substitution of glycine, serine, cysteine,threonine, asparagine, glutamine tyrosine, alanine, valine, isoleucine,leucine, methionine, phenylalanine, lysine, or a D-arginine for arginineat position 36;

[0030] (b) substitution of an oxidation-resistant amino acid fortryptophan at position 31;

[0031] (c) substitution of at least one of: tyrosine for valine atposition 16; lysine for serine at position 18; aspartic acid forglutamic acid at position 21; serine for glycine at position 22;arginine for glutamine at position 23; arginine for alanine at position24; and glutamine for lysine at position 26; and

[0032] (d) substitution of at least one of: glycine, serine, or cysteinefor alanine at position 8; aspartic acid, glycine, serine, cysteine,threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine,leucine, methionine, or phenylalanine for glutamic acid at position 9;serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine,valine, isoleucine, leucine, methionine, or phenylalanine for glycine atposition 10; and glutamic acid for aspartic acid at position 15; and

[0033] (e) substitution of glycine, serine, cysteine, threonine,asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine,methionine, or phenylalanine, or the D- or N-acylated or alkylated formof histidine for histidine at position 7; wherein, in the substitutionsis (a), (b), (d), and (e), the substituted amino acids can optionally bein the D-form and the amino acids substituted at position 7 canoptionally be in the N-acylated or N-alkylated form.

[0034] Because the enzyme, dipeptidyl-peptidase IV (DPP IV), may beresponsible-for the observed rapid in vivo inactivation of administeredGLP-1, [see, e.g., Mentlein, R., et al., Eur. J. Biochem., 214:829-835(1993)], administration of GLP-1 analogs and derivatives that areprotected from the activity of DPP IV is preferred, and theadministration of Gly⁸-GLP-1 (7-36)NH₂, Val⁸-GLP-1 (7-37)OH,a-methyl-Ala⁸-GLP-1 (7-36)NH₂, and Gly⁸-Gln²¹-GLP-1 (7-37)OH, orpharmaceutically-acceptable salts thereof, is more preferred.

[0035] The use in the present invention of a molecule claimed in U.S.Pat. No. 5,188,666, herein incorporated by reference, is preferred. Suchmolecule is selected from the group consisting of a peptide having theamino acid sequence:

His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-X  (SEQID NO:2)

[0036] wherein X is selected from the group consisting of Lys andLys-Gly; and a derivative of said peptide, wherein said peptide isselected from the group consisting of: a pharmaceutically-acceptableacid addition salt of said peptide; a pharmaceutically-acceptablecarboxylate salt of said peptide; a pharmaceutically-acceptable loweralkylester of said peptide; and a pharmaceutically-acceptable amide ofsaid peptide selected-from the group consisting of amide, lower alkylamide, and lower dialkyl amide.

[0037] Another preferred group of molecules for use in the presentinvention consists of compounds disclosed in U.S. Pat. No. 5,512,549,herein incorporated by reference, having the general formula:

R¹-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Xaa-Glu-Phe-Ile-Ala-Trp-

[0038] (SEQ ID NO:3)

[0039] and pharmaceutically-acceptable salts thereof, wherein R¹ isselected from the group consisting of 4-imidazopropionyl,4-imidazoacetyl, or 4-imidazo-a, a dimethyl-acetyl; R² is selected fromthe group consisting of C₆-C₁₀ unbranched acyl, or is absent; R³ isselected from the group consisting of Gly-OH or NH₂; and, Xaa is Lys orArg, may be used in present invention.

[0040] More preferred compounds of SEQ ID NO:3 for use in the presentinvention are those in which Xaa is Arg and R² is C₆-C₁₀ unbranchedacyl.

[0041] Highly preferred compounds of SEQ ID NO:3 for use in the presentinvention are those in which Xaa is Arg, R² is C₆-C₁₀ unbranched acyl,and R is Gly-OH.

[0042] More highly preferred compounds of SEQ ID NO:3 for use in thepresent invention are those in which Xaa is Arg, R² is C₆-C₁₀ unbranchedacyl, R is Gly-OH, and R¹ is 4-imidazopropionyl.

[0043] The most preferred compound of SEQ ID NO:3 for use in the presentinvention is that in which Xaa is Arg, R² is C₈ unbranched acyl, R³ isGly-OH, and R is 4-imidazopropionyl.

[0044] The use of Val⁸-GLP-1 (7-37)OH or a pharmaceutically-acceptablesalt thereof, as claimed in U.S. Pat. No. 5,705,483, herein incorporatedby reference, in the present invention is highly preferred.

[0045] Methods for preparing the GLP-1, GLP-1 analogs, or GLP-1derivatives useful in the present invention are well-known in the artand are easily within the grasp of ordinarily skilled protein chemistsor biochemists. The amino acid portion of the active compound used inthe present invention, or a precursor thereto, can be made either bysolid-phase synthetic chemistry, purification of GLP-1 molecules fromnatural sources, or recombinant DNA technology. Routine syntheticorganic techniques enable the alkylation and acylation of the GLP-1derivatives.

[0046] The term “GLP-1 related-compound” refers to any compound fallingwithin the GLP-1, GLP-1 analog, or GLP-1 derivative definition.

[0047] The term “preservative” refers to a compound added to apharmaceutical formulation to act as an anti-microbial agent. Aparenteral formulation must meet guidelines for preservativeeffectiveness to be a commercially viable multi-use product. Amongpreservatives known in the art as being effective and acceptable inparenteral formulations are benzalkonium chloride, benzethonium,chlorohexidine, phenol, m-cresol, benzyl alcohol, methylparaben,chlorobutanol, o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate,thimerosal, benzoic acid, and various mixtures thereof. See, e.g.,Wallhauser, K., Develop. Biol. Standard, 24: 9-28 (Basel, S. Krager,1974).

[0048] The term “buffer” or “pharmaceutically acceptable buffer” refersto a compound that is known to be safe for use in protein formulationsand that has the effect of controlling the pH of the formulation at thepH desired for the formulation. Pharmaceutically acceptable buffers forcontrolling pH at a moderately acid pH to a moderately basic pH include,for example, such compounds as phosphate, acetate, citrate, TRIS,arginine-, or histidine.

[0049] The term “isotonicity agent” refers to a compound that istolerated physiologically and imparts a suitable tonicity to aformulation to prevent the net flow of water across the cell membrane.Compounds, such as glycerin, are commonly used for such purposes atknown concentrations. Other acceptable isotonicity agents include saltsand sugars, e.g., NaCl, dextrose, mannitol, and lactose. Glycerol at aconcentration of 12 to 25 mg/mL is preferred as an isotonicity agent.

[0050] GLP-1 related compounds described above are administered byinhalation in a dose effective manner to introduce circulatingtherapeutic levels which results in reducing abnormally high bloodglucose levels. Therapeutic serum levels of GLP-1 are in the range of0.1 to about 10.0 ng/ml, preferably about 0.3 to about 5.0 ng/ml andmost preferably about 0.5 to about 3.0 ng/ml. Such administration can beeffective for treating disorders such as diabetes, hyperglycemia, orinsulin resistance. Achieving therapeutically effective doses of GLP-1related compounds requires administering an inhalation dose of about 0.5μg/kg to about 100 μg/kg of the GLP-1 molecule, preferably about 1.0μg/kg to about 50 μg/kg, more preferably about 2.0 μg/kg to about 25μg/kg, and most preferably about 2.5 μg/kg-to about 15 μg/kg. Atherapeutically effective amount can be determined by a knowledgeablepractitioner, who will take into account factors including native GLP-1blood levels, blood glucose levels, the physical condition of thepatient, the patient's pulmonary status, and the like.

[0051] According to the invention, GLP-1 and GLP-1 analogs andderivatives are delivered by inhalation to achieve rapid absorption inthe lungs. Administration by inhalation can result in pharmacokineticscomparable to subcutaneous administration of these substances.Inhalation of GLP-1 and GLP-1 analogs and derivatives leads to a rise inthe level of circulating insulin followed by a rapid fall in bloodglucose levels. Different inhalation devices typically provide similarpharmacokinetics when similar particle sizes and similar levels of lungdeposition are compared.

[0052] According to the invention, GLP-1 and GLP-1 analogs andderivatives can be delivered by any of a variety of inhalation devicesknown in the art for administration of a therapeutic agent byinhalation. These devices include metered dose inhalers, nebulizers, drypowder inhalers, sprayers, and the like. Preferably, GLP-1 and GLP-1analogs and derivatives are delivered by a dry powder inhaler or asprayer. There are a several desirable features of an inhalation devicefor administering GLP-1 and GLP-1 analogs and derivatives. For example,delivery by the inhalation device is advantageously reliable,reproducible, and accurate. The inhalation device should deliver smallparticles, e.g. less than about 10 μm mass median aerodynamic diameter(MMAD), preferably about 1-5 μm MMAD, for good respirability. Somespecific examples of commercially available inhalation devices, or thosein late stage development, suitable for the practice of this inventionare Turbohaler™ (Astra), Rotahaler® (Glaxo), Diskus® (Glaxo), Spiros™inhaler (Dura), devices being developed by Inhale Therapeutics, AERx®(Aradigm), the Ultravent® nebulizer (Mallinckrodt), the Acorn II®nebulizer (Marquest Medical Products), the Ventolin® metered doseinhaler (Glaxo), the Spinhaler® powder inhaler (Fisons), or the like.

[0053] As those skilled in the art will recognize, the formulation ofGLP-1 and GLP-1 analogs and derivatives, the quantity of the formulationdelivered, and the duration of administration of a single dose depend onthe type of inhalation device-employed. For some aerosol deliverysystems, such as nebulizers, the frequency of administration and lengthof time for which the system is activated will depend mainly on theconcentration of the GLP-1 molecule in the aerosol. For example, shorterperiods of administration can be used at higher concentrations of GLP-1and GLP-1 analogs and derivatives in the nebulizer solution. Devicessuch as metered dose inhalers can produce higher aerosol concentrations,and can be operated for shorter periods to deliver the desired amount ofGLP-1 and GLP-1 analogs and derivatives. Devices such as powder inhalersdeliver active agent until a given charge of agent is expelled from thedevice. In-this type of-inhaler, the amount of GLP-1 and GLP-1 analogsand derivatives in a given quantity of the powder determines the dosedelivered in a single administration.

[0054] The particle size of the GLP-1 and GLP-1 analogs and derivativesin the formulation delivered by the inhalation device is critical withrespect to the ability of protein to deposit in the lungs, andpreferably in the lower airways or alveoli. Preferably, the GLP-1 andGLP-1 analogs and derivatives is formulated so that at least about 10%of the peptide delivered is deposited in the lung, preferably about 10%to about 20%, or more. It is known that the maximum efficiency ofpulmonary deposition for mouth breathing humans is obtained withparticle sizes of about 2 μm to about 3 μm MMAD. When particle sizes areabove about 5 μm MMAD, pulmonary deposition decreases substantially.Particle sizes below about 1 μm MMAD cause pulmonary deposition todecrease, and it becomes difficult to deliver particles with sufficientmass to be therapeutically effective. Thus, particles of GLP-1 and GLP-1analogs and derivatives delivered by inhalation have a particle sizepreferably less than about 10 μm MMAD, more preferably in the range ofabout 1 μm to about 5 μm MMAD, and most preferably in the range of about2 μm to about 3 μm MMAD. The formulation of GLP-1 and GLP-1 analogs andderivatives is selected to yield the desired particle size in the choseninhalation device.

[0055] Advantageously for administration as a dry powder, GLP-1 andGLP-1 analogs and derivatives are prepared in a particulate formresulting in an emitted particle size less than about 10 μm MMAD,preferably about 1 to about 5 μm MMAD, and most preferably about 2 μm toabout 3 μm MMAD. The preferred particle size is effective for deliveryto the alveoli of the patient's lung. Preferably, the dry powder islargely composed of particles produced so that a majority of theparticles have a size in the desired range. Advantageously, at leastabout 50% of the dry powder is made of particles having a diameter lessthan about 10 μm MMAD. Such formulations-can be achieved by spraydrying, milling, or critical point condensation of a solution containingthe particular GLP-1 molecule and other desired ingredients. Othermethods also suitable for generating particles useful in the currentinvention are known in the art.

[0056] The particles are usually separated from a dry powder formulationin a container and then transported into the lung of a patient via acarrier air stream. Typically, in current-dry powder inhalers, the forcefor breaking up the solid is provided solely by the patient'sinhalation. One suitable dry powder inhaler is the Turbohaler™manufactured by Astra (Sodertalje, Sweden). In another type of inhaler,air flow generated by the patient's inhalation activates animpeller-motor which deagglomerates the GLP-1 molecule particles. TheDura Spiros™ inhaler is such a device.

[0057] Formulations of GLP-1 and GLP-1 analogs and derivatives foradministration from a dry powder inhaler typically include a finelydivided dry-powder containing peptide, but the powder can also include abulking agent, carrier, excipient, another additive, or the like.Additives can be included in a dry powder formulation of GLP-1 and GLP-1analogs and derivatives-, for example, to dilute the powder as requiredfor delivery from the particular powder inhaler, to facilitateprocessing of the formulation, to provide advantageous powder propertiesto the formulation, to facilitate dispersion of the powder from theinhalation device, to stabilize the formulation (e.g., antioxidants orbuffers), to provide taste to the formulation, or the like.Advantageously, the additive does not adversely affect the patient'sairways. The GLP-1 and GLP-1 analogs and derivatives can be mixed withan additive at a molecular level or the solid formulation can includeparticles of the peptide mixed with or coated on particles of theadditive. Typical additives include mono-, di-, and polysaccharides;sugar alcohols and other polyols, such as, for example, lactose,glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose,mannitol, starch, or combinations thereof; surfactants, such assorbitols, diphosphatidyl choline, or lecithin; or the like. Typicallyan additive, such as a bulking agent, is present in an amount effectivefor a purpose described above, often at about 50% to about 90% by weightof the formulation. Additional agents known in the art for formulatingproteins can also be included in the formulation.

[0058] In another aspect of the invention a spray including GLP-1 andGLP-1 analogs and derivatives can be produced by forcing a suspension orsolution of the peptide through a nozzle under pressure. The nozzle sizeand configuration, the applied pressure, and-the liquid feed rate can bechosen to achieve the desired output and particle size. An electro-spraycan be produced, for example, by an electric field in connection with acapillary or nozzle feed. Advantageously, proplets of GLP-1 and GLP-1analogs and derivatives delivered by a sprayer have an inhaled dropletsize less than about 10 μm MMAD, preferably in the range of about 1 μmto about 5 μm MMAD, and most preferably about 2 μm to about 3 μm MMAD.

[0059] Formulations of GLP-1 and GLP-1 analogs and derivatives suitablefor use with a sprayer typically are about 1 mg to about 20 mg of thepeptide per ml of solution. The formulation can include agents such asan excipient, a buffer, an isotonicity agent, a preservative, asurfactant, and metal cations. The formulation can also include anexcipient or agent to stabilize the peptide such as a buffer, a reducingagent, a bulk protein, or a carbohydrate. Bulk proteins useful informulating GLP-1 and GLP-1 analogs and derivatives include albumin,protamine, or the like. Typical carbohydrates useful in formulatingGLP-1 and GLP-1 analogs and derivatives include sucrose, mannitol,lactose, trehalose, glucose, or the like. Formulations of GLP-1 andGLP-1 analogs and derivatives can also include a surfactant, which canreduce or prevent surface-induced aggregation of the peptide caused byatomization of the solution in forming an aerosol. Various conventionalsurfactants can be employed, such as polyoxyethylene fatty acid estersand alcohols, and polyoxyethylene sorbitol fatty acid esters. Amountswill generally range between 0.001 and 4% by weight of the formulation.Other surfactants such as diphosphatidyl choline or lecithin can also beused. Especially preferred surfactants for purposes of this inventionare polyoxyethylene sorbitan monooleate, polysorbate 80, polysorbate 20,or the like. Additional agents known in the art for formulating proteinscan also be included in the formulation.

[0060] GLP-1 and GLP-1 analogs and derivatives can be administered by anebulizer, such as jet nebulizer or an ultrasonic nebulizer. Typically,in a jet nebulizer, a compressed air source is used to create ahigh-velocity air jet through an orifice. As the gas expands beyond thenozzle, a low-pressure region is created, which draws a solution of thepeptide through a capillary tube connected to a liquid reservoir. Theliquid stream from the capillary tube is sheared into unstable filamentsand droplets as it exits the tube, creating the aerosol. A range ofconfigurations, flow rates, and baffle types can be employed to achievethe desired performance characteristics from a given jet nebulizer. Inan ultrasonic nebulizer, high-frequency electrical energy is used tocreate vibrational, mechanical energy, typically employing apiezoelectric transducer. This energy is transmitted to the peptideformulation either directly or through a coupling fluid, creating anaerosol. Advantageously, droplets of GLP-1 and GLP-1 analogs andderivatives delivered by a nebulizer have a particle size less thanabout 10 μm MMAD, preferably in the range of about 1 μm to about 5 μmMMAD, and most preferably about 2 μm to about 3 μm MMAD.

[0061] Formulations of GLP-1 and GLP-1 analogs and derivatives suitablefor use with a nebulizer, either jet or ultrasonic, typically include anaqueous solution of the peptide at a concentration of about 1 mg toabout 20 mg per ml of solution. The formulation can include agents suchas an excipient, a buffer, an isotonicity agent, a preservative, asurfactant, and a divalent metal cation. The formulation can alsoinclude an excipient or agent to stabilize the peptide, such as abuffer, a reducing agent, a bulk protein, or a carbohydrate. Bulkproteins useful in formulating GLP-1 and GLP-1 analogs and derivativesinclude albumin, protamine, or the like. Typical carbohydrates useful informulating GLP-1 related proteins include sucrose, mannitol, lactose,trehalose, glucose, or the like. Formulations of the GLP-1 and GLP-1analogs and derivatives can also include a surfactant, which can reduceor prevent surface-induced aggregation of the peptide caused byatomization of the solution in forming an aerosol. Variousconventional-surfactants can be employed, such as polyoxyethylene fattyacid esters and alcohols, and polyoxyethylene sorbital fatty acidesters. Amounts will generally range between 0.001 and 4% by weight ofthe formulation. Other surfactants such as phosphatidyl choline orlethicin can also be used. Especially preferred surfactants for purposesof this invention are polyoxyethylene sorbitan monooleate, polysorbate80, polysorbate 20, or the like. Additional agents known in the art forformulation of a protein such as GLP-1 related molecules can also beincluded in the formulation.

[0062] Another aspect of the invention involves a metered dose inhaler(MDI). In this embodiment, a propellant, GLP-1 and GLP-1 analogs andderivatives, and any excipients or other additives are contained in acanister as a mixture including a liquefied compressed gas. Actuation ofthe metering valve releases the mixture as an aerosol, preferablycontaining inhaled particles in the size range of less than about 10 μmMMAD, preferably about 1 μm to about 5 μm MMAD, and most preferablyabout 2 μm to about 3 μm MMAD. The desired aerosol particle size can beobtained by employing a formulation of GLP-1 and GLP-1 analogs andderivatives produced by various methods known to those of skill in theart, including jet-milling, spray drying, critical point condensation,or the like. Preferred metered dose inhalers include those manufacturedby 3M or Glaxo and employing a hydrofluorocarbon propellant.

[0063] Formulations of GLP-1 and GLP-1 analogs and derivatives for usewith a metered-dose inhaler device will generally include a finelydivided powder containing peptide as a suspension in a non aqueousmedium, for example, suspended in a propellant with the aid of asurfactant. The propellant may be any conventional material employed forthis purpose, such as chlorofluorocarbon, a hydrochlorofluorocarbon, ahydrofluorocarbon, or a hydrocarbon, including trichlorofluoromethane,dichlorodifluoromethane, dichlorotetrafluoroethanol and1,1,1,2-tetrafluoroethane, HFA-134a (hydrofluroalkane-134a), HFA-227(hydrofluroalkane-227), or the like. Preferably the propellant is ahydrofluorocarbon. The surfactant can be chosen to stabilize the GLP-1molecule as a suspension in the propellant, to protect the active agentagainst chemical degradation, and the like. Suitable surfactants includesorbitan trioleate, soya lecithin, oleic acid, or the like. In somecases solution aerosols are preferred using solvents such as ethanol.Other surfactants such as diphosphatidyl choline or lethicin can also beused. Additional agents known in the art for formulating proteins canalso be included in the formulation.

[0064] The present invention also relates to a pharmaceuticalcomposition or formulation including GLP-1 and GLP-1 analogs andderivatives and suitable for administration by inhalation. According tothe invention, GLP-1 and GLP-1 analogs and derivatives can be used formanufacturing a formulation or medicament suitable for administration byinhalation. The invention also relates to methods for manufacturingformulations including GLP-1 related molecules in a form that issuitable for administration by inhalation. For example, a dry powderformulation can be manufactured in several ways, using conventionaltechniques. Particles in the size range appropriate for maximaldeposition in the lower respiratory tract can be made by micronizing,milling, spray drying, or the like. And a liquid-formulation can bemanufactured by dissolving the peptide in a suitable solvent, such aswater, at an appropriate pH, including buffers or other excipients.

[0065] The present invention may be better understood with reference tothe following examples. These examples are intended to be representativeof specific embodiments of the invention, and are not intended aslimiting the scope of the invention.

EXAMPLES Serum Pharmacokinetics of Val⁸-GLP-1 in Beagle Dogs FollowingPulmonary Administration

[0066] The GLP-1 analog, Val⁸-GLP-1 (7-37)OH (SEQ ID NO:4) was preparedin E coli using conventional recombinant DNA techniques and purified tohomogeneity.

NH₂-His-Val-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly-OH  (SEQID NO: 4)

[0067] One group of 6 female beagle dogs was exposed to inhaledVal⁸-GLP-1 for 15 minutes at an average aerosol concentration of 77.2μg/L generated from a solution of Val⁸-GLP-1 in sterile water. Theseanimals were dosed with 100 μg/kg via subcutaneous administrationapproximately 1-week after inhalation exposures. Tidal volume, breathingrate, and minute volume were monitored prior to and throughout theexposure period. Blood was collected for analysis of plasma levels ofVal⁸-GLP-1 at various time points following inhalation and subcutaneousadministration. Bronchoalveolar lavage (BAL) fluid was collectedapproximately 4 hours postexposure and analyzed for LDH, total protein,cell counts, and white cell differentials.

[0068] The delivery of Val⁸-GLP-1 was well tolerated with an inhaleddose of 1198 μg/kg and an estimated deposited lung dose of 240 μg/kg.Subcutaneous administration of 100 μg/kg was also well tolerated by allanimals. Inhalation and subcutaneous administration of Val⁸-GLP-1 wasdelivered using formulated and formulated material.

[0069] There were no treatment-related clinical observations; bodyweights were not adversely affected by Val⁸-GLP-1. Only minor lungeffects were observed. Increases in both tidal volume and minute volumewere observed during the 15 minute inhalation exposures but the datawere highly variable. No significant changes were observed for LDH, redblood cell counts, white blood cell counts, neutrophils, lymphocytes,eosinophils, epithelial cells, macrophages, basophils, or monocytes.There was a mild increase in total protein following aerosol delivery ofVal⁸-GLP-1.

[0070] Results from this study demonstrated that there was goodbioavailability of Val⁸-GLP-1 (40%, based on AUC) delivered to the lungsof beagle dogs by inhalation relative to subcutaneous administration.Val⁸-GLP-1 was well tolerated for up to 15 minutes with minimal effectson the lungs at an average inhaled dose of 1198 μg/kg, which was ano-observed-adverse-effects level (NOAEL) in this study.

Preparation of Dose Solutions

[0071] Solutions of Val⁸-GLP-1 were prepared on the days of dosing atconcentrations of 0.5 mg/ml or 8 mg/ml in sterile water for subcutaneousadministration and pulmonary administration, respectively. An additionalsolution of Val⁸-GLP-1 (8 mg/ml) was prepared at the end of live phasein order to determine the particle size distribution of aerosolizedVal⁸-GLP-1. The solutions were filtered through a low protein binding0.22 micron canister filter. The pH of the solution was adjusted withsodium hydroxide solution to 7.47.

Test Animals

[0072] Six females Beagle dogs (Marshall Farms, North Rose, N.Y.) wereused in this study. Each animal was uniquely identified by a five-digitanimal number and a seven-digit tattoo (located on inner ear) numberrecorded on their cage card. All of the animals were acclimated to therestraint slings prior to beginning the study. The weight range of theanimals at the start of the study was 8.4 to 11.1 kg. The age of theanimals at the start of the study was 33 to 37 weeks.

Test Animal Housing and Care

[0073] Animals were pair housed in stainless steel cages except on thedays of exposure. Each animal was individually housed on the day ofexposure in order to monitor their feeding regimen. Rooms werethermostatically set to maintain a temperature of 70° F. and maintainthe actual temperature within ±8° F. from that set point. Theenvironmental control system is designed to maintain a relative humidityof 20% and a maximum of 80%. Light was on a 12-hour cycle, with lightson between 0600 and 1800 hours. Subsequently, lights were off between1800 and 0600 hours except when blood samples were collected. Animalswere fed once daily with Hill's Science Diet. Animals were fasted forapproximately 12-hours prior to exposure. Tap water was provided adlibitum except during exposures.

Treatment Groups and Study Duration

[0074] All 6 dogs were exposed for 15 minutes to aerosolized Val⁸-GLP-1.The targeted deposited lung dose for Val⁸-GLP-1 exposures was 200 μg/kgof body weight. Approximately 7 days after pulmonary administration ofVal⁸-GLP-1, all dogs were dosed with 100 μg/kg Val⁸-GLP-1 of body weightvia subcutaneous administration.

Exposure System

[0075] The dogs were tested while standing in restraint slings. Twolayers of 0.03 inch latex sheets were placed around the animals' necksto form a nonrestrictive airtight seal. A custom built, 11-L head-dome,similar to that described by Allen et al. (J Appl Toxicol 1995;15:13-17) was placed over the dogs' heads and secured to the sling.Airflow was exhaust driven via a transvector located on the exhaust sideof the dome. Because the helmet was airtight and the neck was sealed,this constituted a head-only exposure system. The total flow ratethrough the dome was approximately. 7.5 L/min.

Aerosol Generation

[0076] The aerosols were generated using a Respirgard II nebulizer withan input of approximately 6.5 L/min. The output from the generatorflowed directly into the head-dome.

Atmospheric Concentration Sampling

[0077] All sampling for total gravimetric concentrations was performedwith in-line filter holders containing type A/E glass fiber filters(Gelman Instruments Co., Ann Arbor, Mich.). Filters sampling from thechamber was carried out at a nominal sampling rate of 1 L/min,calibrated with a portable mass airflow calibrator (Model 830, SierraInstruments, Carmel Valley, Calif.). Sampling duration was 15 minutes.Particle size analysis was performed with a Sierra Model 218K AmbientCascade Impactor (Anderson Samplers, Inc, Atlanta, Ga.). Cascadesampling from the chamber was carried out at a nominal sampling rate of3 L/min, calibrated with a portable mass airflow calibrator (Model 830,Sierra Instruments, Carmel Valley, Calif.). Sampling duration was 31minutes. Filters were allowed to dry for approximately 30-minutes beforebeing re-weighed.

Dose Determination

[0078] The dose of Val⁸-GLP-1 inhaled during a 15-minute exposure wasestimated as follows: the mean minute volume (mL) during the 15 minuteexposure was multiplied by the exposure duration to yield the total airbreathed (L) during the inhalation exposure. This value was multipliedby the aerosol concentration (μg/L) to determine total dose (mg). Theinhaled dose (μg/kg) was calculated by dividing the total dose (μg) bythe animal's body weight (kg).

[0079] The dose of Val⁸-GLP-1 deposited in the lungs was estimated asfollows: inhaled dose (μg/kg) was multiplied by 20 percent to yieldestimated deposited lung dose (μg/kg). Aerosols with a MMEAD rangingfrom 1-2 μm MMAD have been shown to deposit in the lung withapproximately a 20 percent efficiency (Schlesinger RB. 1985. Comparativedeposition of inhaled aerosols in experimentalanimals and humans: areview. J Toxicol Environ Health 15:197-214).

Pulmonary Function

[0080] All animals were weighed on Days −5, 0, and 7. Breathing patterns(tidal volume, breathing frequency, and minute volume) were monitoredusing a size ‘0’ pneumotachograph connected to a port on the head-dome.The signals were collected on a personal computer using the Buxco XAData Acquisition System (Buxco Electronics, Inc., Sharon, Conn.). Atleast 15 minutes of preexposure data were collected before the exposuresbegan, followed by data collection throughout the 15-minute exposureperiods. All data were analyzed as 5-minute averages.

Bronchoalveolar Lavage

[0081] Bronchoalveolar lavage (BAL) was performed approximately 4 hoursafter each dosing regimen. The animals were anesthetized with anintravenous injection of 2% Brevital prior to the BAL procedure.Bronchoalveolar lavage was performed using a pediatric fiberopticbronchoscope (Olympus, model BF, Type 3C10, Lake Success, N.Y.). Thebronchoscope tip was wedged in a 5^(th) to 7^(th) generation airway of alower lobe. The BALs were alternated between the right and left lunglobes. Two 10-mL aliquots were instilled and gently suctioned out.Aliquots of the recovered lavage fluid were used to determine totalwhite cell counts, total red-cell counts, white cell differentials,total protein, and lactate dehydrogenase.

Cell Counts and Differentials

[0082] A complete blood count was done on the unconcentrated BAL fluidusing a Technicon H1 System (Technicon Instruments Corporation). Celldifferentials for the BAL were done by microscopic evaluation of 200Wright stained cells.

Blood Collection

[0083] For analysis of plasma pharmacokinetics of Val⁸-GLP-1,approximately 2 to 3 mL of blood were collected in EDTA vacutainer tubesfrom the cephalic or jugular vein prior to exposure and at 0.25, 0.5, 1,2, 3, 4, 6, 8, and 12 hours postexposure. To obtain the plasma, eachtube was centrifuged at approximately 3000 rpm for 10 minutes at 10° C.The plasma samples were stored at −70° C. until sent for assay.

Exposure Concentration/Particle Size and Lung Dose

[0084] The average exposure concentration for each dog exposed toVal⁸-GLP-1 ranged from 64.0 to 101.3 μg/L. The mean (±SD) concentrationfor all animals was 77.2±16.9 μg/L. Particle size was measured as a massmedian equivalent aerodynamic diameter (MMEAD) of 0.91 μm with ageometric standard deviation (GSD) of 2.37.

[0085] The average dose of Val⁸-GLP-1 deposited in lungs of 6 dogstreated with Val⁸-GLP-1 was calculated as 240±42 μg/kg (mean±SD). Themean inhaled dose (that which entered the respiratory tract neglectingdeposition) was 1198±208 μg/kg (mean±SD). Individual animal data areshown in the table below.

Estimated Lung Dose for Animals Exposed to Aerosols of Val⁸-GLP-1

[0086] Aerosol Average Inhaled Deposited Animal Cone. Minute Lung DoseLung Dose Number (μg/L) Volume (ml) (μg/kg) (μg/kg) 27682  0.07717* 84061060 212 27684  0.07717* 9563 1030 206 27685 0.10133 7260 1350 270 276860.06400 12733 1360 272 27687 0.06733 9214 950 190 27689 0.07600 118901400 288

[0087] No significant changes in body weight were observed during thetreatment phase. The initial (Day −5) and final (Day 7) body weightswere 9.5±0.9 (mean±SD; n=6) and 9.7±0.9 kg, respectively. No markedchanges in breathing frequency was observed. Slight increases in tidalvolume and minute volume were measured but the data was highly variablepotentially due to the short acclimation period prior to studyinitiation. Individual animal data are shown in the table below.

Changes in Pulmonary Function During Exposure to Val⁸-GLP-1

[0088] Breathing Animal Tidal Volume (mL) Frequency (bpm) Minute Volume(mL/min) Number 5* 10* 15* 5* 10* 15* 5* 10* 15* 27682 119.0 152.1 105.174 86.0 95.7 7265 9360 8592 27684 104.6 99.4 100.8 119.2 118.2 108.710300 9117 9272 27685 165.9 178.5 209.6 58.1 49.4 42.5 7938 7465 637727686 325.1 586.2 374.4 56.3 33.8 59.4 12200 13300 12700 27687 245.9221.6 183.1 39.5 45.5 50.5 9288 9573 8781 27689 184.5 367.6 454.0 39.846.8 33.3 7271 13400 15000 Mean 190.8 267.6 237.8 64.6 63.3 65.0 904410369 10120 Stdev 82.8 180.7 145.3 29.8 32.2 30.3 1956 2426.1 3141 Mean239.0 216.8 207.6** 45.9 51.4 64.4** 8448 8744.8 8223** (baseline) Stdev161.4 137.2 209.5 12 10.2 32.3 2614 2614 1566 (baseline)

Comparison of Pharmacokinetics Following Subcutaneous and PulmonaryAdministration of Val⁸-GLP-1

[0089] Delivery C_(max) T_(max) AUC_(0-t′(1)) T½(α) Route (ng/mL) (h)(ng*h/mL) (hours) Subcutaneous 10.53 ± 1.06 0.71 ± 0.14 36.37 ± 2.181.26 ± 0.11 Inhalation  8.66 ± 0.90 1.54 ± 0.59 35.21 ± 5.91 1.19 ± 0.11

[0090] Plasma concentrations of immunoreactive Val⁸-GLP-1 (Table 3) weremeasured by a competitive radioimmunoassay (RIA). The absorption ofVal⁸-GLP-1 via both delivery routes appeared to be rapid, reachingsubstantial plasma concentrations at 15 minutes postdose. The plasmatime profiles were similar for the subcutaneous injections andinhalation. The average Tmax value for inhalation was greater than thatfor subcutaneous injection. Also, the elevated plasma Val⁸-GLP-1concentration (close to Cmax) achieved by inhalation appeared to remainnear that level for a longer period of time than following subcutaneousinjection.

[0091] Based on the average AUC values, the bioavailability of inhaledVal⁸-GLP-1 (averaged inhaled dose of 1198 μg/kg) relative tosubcutaneous injection (100 μg/kg) was approximately 7.7%. On the basisof deposited lung dose, estimated as 240 μg/kg, the bioavailabilityrelative to-subcutaneous injection was 40%.

We claim:
 1. A method of administering a glucagon-like peptide-1(GLP-1)molecule comprising, administering an effective amount of a GLP-1molecule selected from the group consisting of GLP-1, GLP-1 analogs, orGLP-1 derivatives to a patient in need thereof by pulmonary means. 2.The method of claim 1, wherein the GLP-1 molecule is delivered to lowerairwaya of the patient.
 3. The method of claim 2, wherein the GLP-1molecule is deposited in the alveoli.
 4. The method of claim-1, whereinthe GLP-1 molecule is inhaled through the mouth of the patient.
 5. Themethod of claim 1, wherein the GLP-1 molecule is administered as apharmaceutical formulation comprising the GLP-1 molecule in apharmaceutically acceptable carrier.
 6. The method of claim 5, whereinthe formulation is selected from the group consisting of a solution inan aqueous medium and a suspension in a non-aqueous medium.
 7. Themethod of claim 6, wherein the formulation is administered as anaerosol.
 8. The method of claim 5, wherein the formulation is in theform of a dry powder.
 9. The method of claim 5, wherein the GLP-1molecule has a particle size of less than about 10 microns MMAD.
 10. Themethod of claim 9, wherein the GLP-1 molecule has a particle size ofabout 1 to about 5 microns MMAD.
 11. The method of claim 10, wherein theGLP-1 molecule has a particle size of about 2 to about 3 microns MMAD.12. The method of claim 1, wherein at least about 10% of the GLP-1molecule delivered is deposited in the lung.
 13. The method of claim 1,wherein the GLP-1 molecule is delivered from an inhalation devicesuitable for pulmonary administration and capable of depositing theGLP-1 molecule in the lungs of the patient.
 14. The method of claim 13,wherein the device is selected from the group consisting of a nebulizer,a metered-dose inhaler, a dry powder inhaler, and a sprayer.
 15. Themethod of claim 14, wherein the device is a dry powder inhaler.
 16. Themethod of claim 1 wherein the GLP-1 molecule is selected from the groupconsisting of GLP-1 analogs and GLP-1 derivatives.
 17. The method ofclaim 16 wherein the GLP-1 molecule is a GLP-1 analog.
 18. The method ofclaim 17 wherein the GLP-1 analog is selected from the group consistingof Val⁸-GLP-1(7-37)OH, Gly⁸-GLP-1 (7-37)OH, and Asp⁸-GLP-1 (7-37)OH. 19.The method of claim 18, wherein the GLP-1 analog is Val⁸-GLP-1 (7-37)OH.
 20. The method of claim 18, wherein the GLP-1 analog is Gly⁸-GLP-1(7-37)OH.
 21. A method for treating diabetes comprising, administeringan effective dose of a GLP-1 molecule to a patient in need thereof bypulmonary delivery.
 22. The method of claim 21, wherein the GLP-1molecule is administered as a pharmaceutical formulation comprising theGLP-1 molecule in a pharmaceutically acceptable carrier.
 23. The methodof claim 21, wherein the GLP-1 molecule is Val⁸-GLP-1 (7-37)OH.
 24. Themethod of claim 21, wherein the GLP-1 molecule is Gly⁸-GLP-1 (7-37)OH.25. The method of claim 21, wherein the GLP-1 molecule is delivered froman inhalation device suitable for pulmonary administration and capableof depositing the GLP-1 molecule in the lungs of the patient.
 26. Themethod of claim 25, wherein the device is a sprayer or a dry powderinhaler.
 27. The method of claim 25, wherein an actuation of the deviceadministers about 40 μg to about 4,000 μg of a GLP-1 molecule.
 28. Themethod of claim 25, wherein an actuation of the device administers about80 μg to about 2,000 μg of a GLP-1 molecule.
 29. The method of claim 25,wherein an actuation of the device administers about 160 μg to about1,000 μg of a GLP-1 molecule.
 30. The method of claim 25, wherein anactuation of the device administers about 320 μg to about 500 μg of aGLP-1 molecule.
 31. A method for treating hyperglycemia comprising,administering an effective dose of a GLP-1 molecule to a patient in needthereof by pulmonary means.
 32. The method of claim 31, wherein theGLP-1 molecule is administered as a pharmaceutical formulationcomprising the GLP-1 molecule in a pharmaceutically acceptable carrier.33. The method of claim 31, wherein the GLP-1 molecule is Val⁸-GLP-1(7-37)OH.
 34. The method of claim 31, wherein the GLP-1 molecule isGly⁸-GLP-1 (7-37) OH.
 35. The method of claim 31, wherein the GLP-1molecule is delivered from an inhalation device suitable for pulmonaryadministration and capable of depositing the GLP-1 molecule in the lungsof the patient.
 36. The method of claim 35, wherein the device isselected from the group consisting of a sprayer and a dry powderinhaler.
 37. The method of claim 35, wherein an actuation of the deviceadministers about 40 μg to about 4,000 μg of GLP-1 molecule.
 38. Themethod of claim 35, wherein an actuation of the device administers about8.0 μg to about 2,000 μg of the GLP-1 molecule.
 39. The method of claim35, wherein an actuation of the device administers about 160 μg to about1,000 μg of GLP-1 molecule.
 40. The method of claim 35, wherein anactuation of the device administers about 320 μg to about 500 μg of theGLP-1 molecule.